| NAC transcription factors are a class of widely found in terrestrial plants,participate in the growth and development of plants,biotic and abiotic stress family.Abiotic stresses including drought,high salt,low temperature and other external environmental factors as well,such as abscisic acid and methyl jasmonate treatment plant hormones,the increased expression of the NAC gene,associated with the expression of the NAC gene was also increased,so increased resistance of plants,in exceptional circumstances can grow normally.In this experiment,the NAC transcription factor Salvia miltiorrhiza,SmNAC1 for the study,obtained by 3’ RACE and RT-PCR method for the full length;yeast two-hybrid method to validate their self-activating transcription activity;and the use of real time-PCR methods for Salvia spatial-temporal expression analysis;transgenic tobacco under high salt,drought and ethylene stress conditions,development of tobacco roots of transgenic tobacco and phenotype control wild-type tobacco,and SmN AC1 gene and marker gene in transgenic tobacco plants and wild-type tobacco expression.Laboratory of previous work using the yeast two-hybrid approach,get some NAC similar sequence in Salvia miltiorrhiza,after 3 ’RACE and RT-PCR method to obtain the full length of the NAC is 1 734 bp,named SmNAC1 gene,encoding 577 amino acidsBioinform atics analysis using online biology software Salvia miltiorrhiza SmNAC1 genes were transeription factors,theoretical isoelectric point of 4.77 and a relative protein component 64,223.42.Conserved regions are 14-1 64 A A,protein secondary strueture analysis,a-helix(H)accounted for 55.5%,β-fold(E)accoulted for 59.6%,β-angle(T)accounted for 13.5%.SmNAC1 not secreted protein;protein localization prediction signal,the endoplasmic reticulum results showed the highest score of 0.850,the results indicate,SmNAC1 positioning endoplasmic reticulum may use predicted transmembrane domain 557 to 575 AA as a significant cross membrane area.Homologous sequences and phylogenetic analysis,SmNA Cl gene belongs to a large group of Part Ⅰ,Section Ⅶ(NAC2 family)subfamily.Using the yeast two-hybrid method validation Salvia miltiorrhiza transcription factor Sm NAC1 self-activated transcriptional activi ty.The use of real time PCR method,the Salvia miltiorrhiza SmNA Cl temporal gene expression analysis showed that,with a three-native wild Salvia miltiorrhiza,Sm NAC1 highest expression of genes in leaves,the expression of differences in roots,stems and flowers unlikely.Using a variety of simulated changes in wildtype Salvia miltiorrhiza stress treatment plants verify that the transcription factor SmNACl expressed in roots and leaves.After ABA stress induced differential expression roots too large,showing an overall increasing trend,but differentially expressed in leaves is not obvious;mannitol and NaCl stress-induced increases in root was evident,but not differentially expressed in leaves obvious;after MeJA stress induced roots increased significantly,leaves showed no significant differences;after ethylene stress-induced,salvia root and leaves an increasing trend are particularly significant;after SA stress-induced,salvia root and leaf expression no significant differences.Salvia miltiorrhiza SmNAC1 will build on pCAM BIA1 305.1 binary expression vector,transformed into the tobacco W38.After positive identification of the plant after determining the success the transgenic.After high salt stress,after 0.1 M NaCl stress,tobacco root growth phenotype of transgenic tobacco displays well-developed.Separate detection of transgenic tobacco SmNAC1 exogenous gene,discovered the trend is to decrease the elevated the transgenic Ⅲ and IV,the same trend;and salt stress marker gene NtSAM1 turn genotype significantly increased expression in tobacco,Description Salvia miltiorrhiza SmNAC1 transcription factors with salt tolerance.Under drought stress,stress treatment after 400mM mannitol,tobacco root growth phenotype of transgenic tobacco displays well-developed.the transgenic tobacco Ⅲ and IV,elevated gene expression SmNAC1 changed significantly,transgenic tobacco under drought stress marker gene NtERD1 OC signifieantly inereased,description,Salvia miltiorrhiza Sm NAC1 transcription factors with drought tolerance.1mM ethylene under stress conditions,the development of roots of tobacco apparent phenotype,normal growth of transgenic,wild-type yellow tobacco has withered,along with the expression SmNA Cl first time point is lowered and then raised,either the transgenic type Ⅲ or IV,elevated both trends are very clear marker gene AOC2 ethylene under stress gene expression was significantly increased,indicating,Salvia SmNAC1 transcription factors may be involved in the ethylene signaling pathway. |
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